大豆种子成熟蛋白PM40基因表达方式分析及其启动子预测

Analysis of Expression Patterns of PM40 Gene in Soybean and Its Promoter Prediction

研究大豆种子成熟蛋白PM40基因在不同逆境条件下及大豆各组织中的表达情况,并对其上游启动子序列进行生物信息学预测,为揭示PM40基因表达本质及其启动子的功能研究、利用提供理论依据。利用qRT-PCR方法,检测PM40基因在干旱、盐、低温、脱落酸条件下的表达;比较该基因在大豆根、茎、叶、花、30DAF、60DAF和90DAF中的表达;克隆PM40基因5′端上游启动子序列,并预测其含有的顺式作用元件。结果表明,PM40基因在干旱、低温和脱落酸处理不同时间下,与未处理相比,表达量均下降,在盐处理条件下,只有处理2 h时表达升高,其余处理时间基因表达下降,说明PM40基因受干旱、低温和脱落酸诱导表达下降,在盐诱导2 h时表达升高;PM40基因在大豆种子中的表达相对较高,尤其是60DAF和90DAF的种子中,相对于根的表达量为35.76,239.75倍;以大豆基因组DNA为模板,克隆PM40基因5′端上游启动子序列1 581 bp,生物信息学预测表明,PM40基因启动子序列中包含多种与种子高表达相关的顺式元件。推测大豆PM40基因启动子可能具有种子特异表达特性。

The expression of soybean PM40 gene under different stress conditions and in soybean tissues were studied,and the upstream promoter sequence was predicted by bioinformatics,which provide the theoretical foundation for further understanding the regulation of PM40 gene expression and utilizing the promoter.The expression patterns of PM40 gene under drought,salt,low temperature and ABA were detected,the expressions of PM40 gene were compared between roots,stems,leaves,flowers,30DAF,60DAF and 90DAF by qRT-PCR.The 5′-flanking upstream sequence of soybean PM40 gene was isolated and analyzed the cis-elements by bioinformatics.The results showed that compared with the untreated,the expressing quantity of PM40 gene decreased under drought,low temperature and ABA treatment at different times,but elevated in the treatment of 2 hours and fell slightly in the rest of treatment time under salt treatment,which illustrated that PM40 gene were down-regulated induced by drought,low temperature and ABA,but up-regulated at 2 hours and down-regulated at the other treatment times induced by salt.The expression of PM40 gene in seeds were relatively high,especially in 60DAF and 90DAF,the relative expression of PM40 gene to roots were 35.76 and 239.75 times.Using soybean genomic DNA as a template,1581 bp upstream promoter sequence of PM40 gene at 5′end was cloned.Bioinformatics prediction showed that PM40 gene promoter sequence contained a variety of cis-elements related to high expression in seeds,and speculated that PM40 gene promoter might have seed-specific expression characteristics.